Method for staining corneocites, method for preparing corneocites specimen and skin analysis system

ABSTRACT

A method for preparing corneocites specimen, which comprises stripping off corneocites from the surface of a skin using an adhesive material, staining the corneocites in a solution of stain in solvent containing a water-miscible organic solvent, and mounting the stained corneocites into an oil and fat constituent and/or composition that is liquid at 1 atm, 25° C.

BACKGROUND OF THE INVENTION

[0001] The present invention relates to a method for stainingcorneocites as well as a method for preparing corneocites specimen whichcan provide useful information for analysis of skin condition andappropriate selection of cosmetic products.

[0002] The present invention also relates to a skin analysis systemwhich can provide useful information for analysis of skin condition andappropriate selection of cosmetic products.

[0003] Skin conditions may constantly change depending on variablefactors including seasonal changes, physical conditions, skin treatment(e.g., application of cosmetic products) or the like. It is thus veryimportant to select an appropriate cosmetic product according to suchvariable factors in order to prevent aging or keep skin in a goodcondition. For this purpose, methods for objectively and scientificallyanalyze skin type and/or condition have been developed. One example ofsuch method involves collecting corneocites from, for example, thesurface of face using an adhesive tape or disc, staining thecorneocites, and analysis skin type and condition using as indicatorsvarious factors such as the arrangement regularity of the corneocites, ashape or size of a horny cell (corneocite), a roughness of the surfaceof horny stratum (keratinized layer), and the presence or absence of anucleus in the horny cell. In such analysis, a staining method usinggentian violet and brilliant green has been used since it can preferablyprovide a high contrast between cytoplasm and its background and thusconfirm both the shape of individual corneocites and the presence orabsence of a nucleus in each horny cell. This method, however, requiresa relatively longer period of time for the staining step and thus cannotimmediately provide data for instant analysis (e.g., a personalconsultation in store). Therefore, conventional methods for selectingcosmetic products based on the analysis of skin using as an indicatorthe shape of horny cell took several days until it could providecosmetic samples after skin analysis, which disadvantageously produced atime lag. Although the amount of dye can be increased in such a case, itmay yet take ten minutes or more for both gentian violet and brilliantgreen to stain cells even when their maximal water-solubility limits(gentian violet=1%, brilliant green=0.5%) are used. Therefore,conventional method still had a disadvantage in terms of staining time.Conventionally, organic solvents have not been used in such stainingprocess since they may reduce staining-specificity or cause degenerationor alteration of cells. Accordingly, a method for staining a horny cellhas not been known yet in which a sample containing corneocites obtainedby stripping off from a skin using an adhesive material (e.g., astripper) is stained wherein a stain solution used in the staining stepcomprises a water-miscible organic solvent.

[0004] Generally, a stained horny cell sample may be mounted to obtainuniform refractive index through the entire sample. Balsam andultraviolet-curing resins have conventionally been used for mountion.However, those mounting materials are not preferable since they mayoften cause color running, which may result in a significant error inautomatic computation of, for example, the size of cell by binarization(by using a binary image of the corneocites). Moreover, an adhesive tapecannot be used to obtain corneocites since the adhesive or tapecomponent thereof may be decomposed by the organic solvent used formountion. For quick analysis of corneocites in, for example, a personalconsultation at a customer desk as described above, automaticcomputation of the size of horny cell by using the binary image of thehorny cell is inevitable and thus a solution to such regarding colorrunning problem has strongly been demanded. Oil and fat constituentand/or composition that is liquid at 1 atm, 25° C. has not been used forsuch mountion. This is because that it is difficult to preserve such aliquid mounting agent for 1 month or more and that such constituents orcompositions have never been used for mountion so far. Accordingly, itis a novel finding that, by using such mounting agents, a corneocitessample can be prepared which allows for an automatic calculation of thesize of a horny cell by using the binary image of the corneocites withlittle or no color running.

[0005] Skin conditions may constantly change depending on variablefactors including seasonal changes, physical conditions, skin treatment(e.g., application of cosmetic products) or the like. It is thus veryimportant to select an appropriate cosmetic product according to suchvariable factors in order to prevent aging or keep skin in a goodcondition. For this purpose, methods for objectively and scientificallyanalyze skin type and/or condition have been developed.

[0006] In order to select cosmetic products suitable for a particularcustomer, for example, conventional methods involved collectingcorneocites from, for example, the surface of face using an adhesivetape or disc with adhesive material, staining the corneocites, andanalysis skin type and condition using as indicators various factorssuch as the size of horny cell, the roughness of the surface of hornystratum (keratinized layer) and the presence or absence of a nucleus inthe horny cell.

[0007] In such analysis, a staining method using gentian violet andbrilliant green has been used since it can preferably provide a highcontrast between cytoplasm and its background and thus confirm both theshape of individual corneocites and the presence or absence of a nucleusin each horny cell.

[0008] This method, however, requires a relatively longer period of timefor the staining step and thus cannot immediately provide data forinstant analysis (e.g., a personal consultation in store). Therefore,conventional methods for selecting cosmetic products based on theanalysis of skin using as an indicator the shape of horny cell tookseveral days until it could provide cosmetic samples after skinanalysis, which disadvantageously produced a time lag.

[0009] Therefore, no sales method has been introduced which cancomplete, within a day (or rather instantly), the whole processcomprising analysis the skin of a customer at a store counter andrecommending the customer a suitable trial cosmetic kit based on theanalysis results so that the customer can purchase and try the kit andthen select and purchase a full-size cosmetic product or products of thesame version contained in the trial kit if he/she likes it or them.

SUMMARY OF THE INVENTION

[0010] One object of the present invention is to provide a method forstaining corneocites, which can stain corneocites quickly and clearly aswell as a method for preparing a corneocites specimen which canimmediately provide useful data for consultation.

[0011] Another object of the present invention is to provide a skinanalysis system, which can stain the corneocites quickly and clearly andprovide useful data for consultation.

[0012] For these purposes, the present inventors worked hard to developa method for staining corneocites, which can stain the corneocitesquickly and clearly as well as a method for preparing a corneocitesspecimen which can immediately provide useful data for consultation. Andthey finally found that the corneocites can be stained quickly andclearly by a method for preparing a sample containing corneocites whichhave been striped off from the surface of the skin of the subject usingan adhesive material and stained in a solution of stain in solventcontaining a water-miscible organic solvent. They also found that colorrunning can be reduced or eliminated by mounting the stained sample inan oil and fat constituent and/or composition that is liquid at 1 atm,25° C. to prepare a horny cell specimen. This may allow for binarizationof horny cell image (producing a monochromic image of the corneocites)and thus enable an automatic computation of the size of the horny cellin the corneocites specimen. Based on these findings, the presentinventors developed the present invention.

[0013] In summary, the present invention relates to the followingtechniques:

[0014] (1) a method for staining a sample containing corneocites whichhave been stripped off from a surface of a skin of a subject using anadhesive material wherein the sample may be stained in a solution ofstain in solvent containing a water-miscible organic solvent;

[0015] (2) a method according to (1) wherein the dye comprises gentianviolet or brilliant green;

[0016] (3) a method according to (1) or (2) wherein the water-miscibleorganic solvent comprises ethanol;

[0017] (4) a method according to any one of (1)-(3) wherein the adhesivematerial comprises an adhesive tape;

[0018] (5) a method for preparing a specimen comprising a stained samplecontaining corneocites which have been stripped off from a surface of askin of a subject using an adhesive material wherein the stainedspecimen is mounted in an oil and fat constituent and/or compositionthat is liquid at 1 atm, 25° C.;

[0019] (6) a method according to (5) wherein the oil and fat constituentand/or composition that is liquid at 1 atm, 25° C. is one or moreselected from the group consisting of silicone oil, fatty acidtriglyceride, ester of higher alcohol and fatty acid, and carbohydrate;

[0020] (7) a method according to (5) or (6) wherein the samplecontaining corneocites is stained according to any one of stainingmethods (1)-(4);

[0021] (8) a method according to any one of (5)-(7) which may be used todetermine or measure one or more selected from a group consisting of asize of a horny cell, presence or absence of nuclear cell, the frequencyof nuclear cell appearance, the arrangement regularity or shape of hornycell and the roughness of the surface of keratinized layer.

[0022] The present invention also relates to a skin analysis systemwhich comprises:

[0023] a staining apparatus for staining a sample containing corneociteswhich haven been stripped off from the surface of the subject's skinusing an adhesive material and stained in a solution of stain in solventcontaining a water-miscible organic solvent;

[0024] a microscope for obtaining an enlarged image of a corneocitesspecimen prepared from the sample stained in the staining apparatus;

[0025] a corneocites image data generating portion for generatingcorneocites image data that is an image data of the enlarged image ofthe corneocites specimen obtained by the microscope;

[0026] a measurement portion for measuring a size of a horny cell usingthe enlarged image of the corneocites specimen;

[0027] a transmitting portion for transmitting to a server via a networktransmits the corneocites image data and information representing thehorny cell size obtained by the measurement portion;

[0028] a receiving portion for receiving from the server via the networkevaluation information representing results obtained by evaluating atleast one of a roughness of a surface of a keratinized layer and acompleteness of the keratinized layer based on the corneocites image andthe horny cell size information; and

[0029] a display control portion which displays the evaluationinformation on a display portion.

[0030] According to the skin analysis system according to the presentinvention, it takes a shorter period of time to stain corneocites whencompared to conventional systems. The inventive system can thereforereduce the time required until receiving, from a server, results(analysis results) obtained by evaluating at least one of the roughnessof the surface of the keratinized layer and the completeness of thekeratinized layer which will then be used in a personal consultation orthe like.

[0031] The skin analysis system according to the present invention maybe preferably designed so that a microscope may provide an enlargedimage of a corneocites specimen which comprises a stained sample mountedin an oil and fat constituent and/or composition that is liquid at 1atm, 25° C. In this way, color-running may be reduced or eliminated, andan enlarged image of corneocites can be thus obtained which can besuitably used to analyze the roughness of the surface of the keratinizedlayer and the completeness of the keratinized layer.

[0032] The skin analysis system according to the present invention maybe preferably designed to further comprise a texture image datagenerating portion for generating texture image data that is a imagedata of an enlarged image of a surface of the subject's skin obtained byan image pickup device, wherein:

[0033] the transmitting portion may transmit the texture image data tothe server;

[0034] the receiving portion may receive from the server the evaluationinformation including evaluation results about a condition of thetexture based on the texture image; and

[0035] the display control portion may display the evaluationinformation on a display portion.

[0036] The skin analysis system according to the present invention maybe preferably designed to further comprise a dark spot image generatingportion which generates B channel image data obtained by extracting Bchannel alone from an enlarged color image of the surface of thesubject's skin obtained by an image pickup device which can be used toanalyze any dark spot on the subject's skin, wherein:

[0037] the transmitting portion may transmit the B channel image data tothe server;

[0038] the receiving portion may receive from the server the evaluationinformation including evaluation results obtained by evaluating any darkspot based on the B channel image data; and

[0039] the display control portion may display the evaluationinformation on a display portion.

[0040] Use of B channel image can eliminate the staining step forstaining the corneocites sample that was conventionally performed fordark spot analysis, which may in turn reduce the time required until theresults of the evaluation obtained by dark spot analysis transmittedfrom the server is displayed on a display portion.

[0041] The skin analysis system according to the present invention maypreferably be designed so that the display control portion may displayanalysis results of the subject's skin obtained on the basis of theevaluation information and analysis results obtained about persons ofthe same generation of the subject on a display portion so that theformer can be compared with the latter. By this configuration, it may bepossible to provide the subject with more useful information than asystem without displaying such reference data.

[0042] The skin analysis system according to the present invention maypreferably be designed so that the display control portion may displayanalysis results of the subject's skin obtained on the basis of theevaluation information and analysis results obtained about persons ofthe same generation having an age equal to the subject's age or 1 or 2years older and younger than the subject's age on a display portion sothat the former can be compared with the latter.

[0043] Further, the display control portion may preferably be designedto display the results obtained by skin analysis of the subject relativeto analysis results obtained about the persons of the same generation ofthe subject. Preferably, the reference data may represent resultsobtained during the same month as the date of current skin analysis. Inthis way, reference data which has taken into account skin conditionvariable according to seasonal changes can be provided.

[0044] The present invention also provides a display data generatingapparatus for skin analysis, which comprises:

[0045] a receiving portion which receives via a network data to be usedin skin analysis of a subject;

[0046] a data generating portion which generates, when evaluationresults of the skin analysis of the subject are obtained on the basis ofthe skin analysis data, a display data in order to display theevaluation results in such a form that the evaluation results can becompared with evaluation results about persons of the same generationhaving an age equal to the subject's age or 1 or 2 years older andyounger than the subject's age; and

[0047] a display transmitting portion which transmits via the networkthe display data to an apparatus having a display device for reproducingan image or video based on the display data.

[0048] The display data generating apparatus according to the presentinvention for displaying results of skin analysis may preferably bedesigned such that the data generating portion may generate a displaydata to be used to display evaluation results of the skin analysis ofthe subject relative to the evaluation results of the persons of thesame generation.

BRIEF DESCRIPTION OF THE DRAWINGS

[0049]FIG. 1 shows a microscopic photograph of a horny cell specimenprepared by the method according to Example 1 of the present invention;

[0050]FIG. 2 shows a microscopic photograph of horny cell specimenprepared by the method according to Comparative Example 1 described inExample 1 of the present invention;

[0051]FIG. 3 shows a microscopic photograph of horny cell specimenprepared by the method according to Comparative Example 2 described inExample 1 of the present invention;

[0052]FIG. 4 is a diagram showing one example of the skin analysissystem according to the present invention;

[0053]FIG. 5 is a block diagram showing a plurality of web sites to beprovided to a customer;

[0054]FIG. 6 is an explanatory diagram of a screen representationdisplayed on a web page;

[0055]FIG. 7 shows an exemplary display displayed on a top (title) webpage of an advice site;

[0056]FIG. 8 shows one example of a screen representation displayed on aweb page titled “1. total skin score (total score)”;

[0057]FIG. 9 shows a first example of a screen representation displayedon a web page titled “2. general chart”;

[0058]FIG. 10 shows a second example of a screen representationdisplayed on a web page titled “2. general chart”;

[0059]FIG. 11 shows an example of a screen representation displayed on aweb page titled “3. skin positioning”;

[0060]FIG. 12 shows an example of a screen representation displayed on aweb page titled “4. roughness of the surface of keratinized layer (theability of corneocites to keep moisture)”;

[0061]FIG. 13 shows an example of a screen representation displayed on aweb page titled “5. completeness of keratinized layer”;

[0062]FIG. 14 shows an example of a screen representation displayed on aweb page titled “6. texture”;

[0063]FIG. 15 shows an example of a screen representation displayed on aweb page titled “7. dark spot and colorevenness”;

[0064]FIG. 16 shows an example of a screen representation displayed on aweb page titled “8. Sensitive skin type”;

[0065]FIG. 17 shows an example of a screen representation displayed on aweb page titled “9. skincare advice”;

[0066]FIG. 18 shows an example of a screen representation displayed on aweb page which presents cosmetic products to be used in “lotion” and“moisture essence” steps;

[0067]FIG. 19 shows an example of a screen representation displayed on aweb page which presents cosmetic products to be used in “cleansing” and“(facial) wash” steps;

[0068]FIG. 20 shows an example of a screen representation displayed on aweb page which presents “special items”;

[0069]FIG. 21 shows an example of a screen representation displayed on aweb page titled “10. trial kit”;

[0070]FIG. 22 shows an example of a screen representation displayed on aweb page titled “11. skin biorhythm”;

DETAIL DESCRIPTION OF THE INVENTION

[0071] Hereinafter, the present invention will be described in moredetail.

[0072] [1] Method for Staining Horny Cell According to the PresentInvention

[0073] A method for staining horny cell according to the presentinvention is characterized by that a sample containing corneocites whichhave been stripped off from a skin using an adhesive material is stainedwith a solution of stain in solvent containing a water-miscible organicsolvent. Examples of adhesive material preferably include adhesivetransparent tapes such as cellophane-tape and adhesive discs such asthose comprising a polyethylene terephthalate plate coated with anadhesive. Among all, adhesive tapes are particularly preferable sincethose can be commercially obtained very easily. A horny cell sampleobtained by stripping corneocites off from the surface of skin usingsuch an adhesive material may be stained directly or transferred toanother adhesive material for use. Preferably, the sample obtained maybe stained directly so that the time required for the process can bereduced. According to the inventive staining method, samples collectedcan be directly stained without affecting the adhesive or the substrateof the adhesive material (e.g., an adhesive tape or disc) used. Anyconventionally-used dye can be used in the inventive staining method,preferably including eosin, hematoxylin, gentian violet, brilliant greenand malachite green, which can be used alone or in combination. Amongall, a combination of gentian violet and brilliant green may beparticularly preferable. This is because that cytoplasm can be stainedwith gentian violet to obtain a high contrast between the cytoplasm andthe background thereby clearly visualizing the shape of corneocites, andthat the stained cytoplasm may in turn act as the background so thatnuclei of the corneocites can be clearly stained with brilliant green.The inventive staining method is characterized by that a sample isstained in a solution of stain in liquid solvent containing awater-soluble organic solvent. Stain may preferably be present in thedyeing solution at an amount of 2-7% by weight. For a combination ofgentian violet and brilliant green, gentian violet may preferably bepresent at an amount of 1.5-5% by weight and more preferably 2-4% byweight while brilliant green may preferably be present at an amount of0.7-2% by weight and more preferably at 0.8-1.5% by weight. Examples ofwater-soluble organic solvent preferably include: alcohol such asmethanol, ethanol, isopropanol or 1,3-butanediol; keton such as acetoneor methyl ethyl ketone; nitrile such as acetinitrile; and ether such astetrahydrofuran. Among all, alcohol is preferable and ethanol isparticularly preferable. Water-soluble organic solvent may preferably bepresent in the liquid solvent at an amount of 3-55% by weight and morepreferably 5-50% by weight. This is because that a larger amount oforganic solvent may not contribute to reduction in the time required forstaining or rather may degrade the adhesive or substrate of adhesivematerial while a smaller amount cannot stain a sample clearly. Underthose conditions, it may take 1-5 minutes to stain a sample, which ismuch shorter than the time required for staining according toconventional methods (10-30 minutes) (using 0.5-1% by weight of gentianviolet and 0.2-0.5% by weight of brilliant green). Further, a very highcontrast can be obtained. Besides, the staining method using thoseconditions can be performed at room temperature.

[0074] [2] Method for Preparing Horny Cell Specimen According to thePresent Invention

[0075] The method for preparing horny cell specimen according to thepresent invention is a method for preparing a specimen comprising astained sample containing corneocites which have been stripped off fromthe surface of skin using an adhesive material, characterized by thatthe stained sample is then mounted in an oil and fat component and/orcomposition that is liquid at 1 atm, 25° C. Examples of stained samplecontaining corneocites which have been stripped off from the surface ofskin using an adhesive material generally include any stained sampleswhich have been prepared as described above. Particularly preferable arethose obtained by the method for staining horny cell according to thepresent invention described in (1) above. Oil and fat constituent and/orcomposition that is liquid at 1 atm, 25° C. to be used in the inventivestaining method may include oil and fat constituents which areconventionally used in cosmetic products and drugs as well ascompositions comprising any combination thereof which are liquid at 1atm, 25° C. Preferable examples may include one or any combination oftwo or more selected from the group consisting of silicone oil, fattyacid triglyceride, ester of higher alcohol and fatty acid, andcarbohydrate which are liquid at 1 atm, 25° C. Generally, mountion in aliquid component may not provide a stability of 1 month or longer.However, this may not be critical for an instant consultation at a storecounter, which is the purpose of the invention. Such oil base may notcause color running and thus a microscopic image of a specimen obtainedcan be processed into a binary image (binarization) based on whether thecorneocites have been stained or not to provide an accurate shape(contour) of horny cell. Then, the size of horny cell can automaticallybe computed accurately. Additional information which can be obtainedfrom a specimen prepared by the method for preparing specimen accordingto the present invention include the presence or absence of nuclearcell, the frequency of nuclear cell appearance, the roughness of thesurface of keratinized layer, the arrangement regularity of corneocites,the shape of horny cell and the like. Specimen according to the presentinvention can preferably be used to determine numerical values for thoseparameters. In summary, horny cell specimens prepared by the inventivemethod may preferably be used to determine one or more selected from thegroup consisting of the size of horny cell, the presence or absence ofnuclear cell, the frequency of nuclear cell appearance, the arrangementregularity of corneocites, the shape of horny cell and the roughness ofthe surface of keratinized layer.

EXAMPLES

[0076] The present invention will hereinafter be described in moredetail in reference to Examples though it should be noted that thepresent invention might not be limited to these examples.

Example 1

[0077] Three horny cell samples were prepared from one panelist bystripping corneocites off from the surface of his/her skin using anadhesive tape, and two of them-were stained according to the inventivestaining method. Particularly, the two samples were immersed in a dyeingsolution of 3 wt % gentian violet and 1 wt % brilliant green in anaqueous solution of 20% ethanol for 2 minutes and then washed in watersufficiently. One of them was mounted in an ultraviolet-curing resin andthen cured by exposure to UV light (Comparative Example 1) while theother was mounted in dimethicone (silicone) according to the presentinvention (Example 1). The remaining sample was immersed in a dyeingsolution of 1 wt % gentian violet and 0.5 wt % brilliant green in waterfor 10 minutes, washed in water sufficiently, and then mounted indimethicone (Comparative Example 2). Microscopic photographs of thesesamples are shown in FIGS. 1-3. As shown in FIGS. 1-3, a low contrastwas obtained due to color running in Comparative Example 1 and thecorneocites were less stained in Comparative Example 2 while thecorneocites were well stained without color running (i.e., a highercontrast was obtained than that obtained in Comparative Example 1).Those results show that a horny cell specimen obtained by the inventivemethod can provide clear and distinct visualization of corneocites.

Example 2

[0078] Average size was calculated for each of the horny cell specimensobtained by Example 1, Comparative Examples 1 and 2. The image of thehorny cell specimen of Example 1 was manually binarized and used tocalculate an average size (μm²). Twenty cells were averaged for eachexample. The results are shown in Table 1 below. Those results show thatautomatic calculation of size gave the same result as the resultobtained by manual calculation according to the present invention. Onthe other hand, automatic calculation of size gave higher values thanthe actual results for the specimens with less clear staining or colorrunning. TABLE 1 SAMPLE AVERAGE SIZE ± VARIANCE AUTOMATIC DETERMINATION568 ± 121 (EXAMPLE 1) AUTOMATIC DETERMINATION 721 ± 237 (COMPARATIVEEXAMPLE 1) AUTOMATIC DETERMINATION 736 ± 175 (COMPARATIVE EXAMPLE 2)MANUAL DETERMINATION 559 ± 134 (EXAMPLE 1)

Example 3

[0079] Similar process was repeated as in Example 1 except for usingvariable amounts of gentian violet. Again, 1% by weight of brilliantgreen was used. Dimethicone was used for mountion. The clearness ofstaining were indicated as: ◯=clear; Δ=less clear; X not clear. Theresults are shown in Table 2 below. Those results show that gentianviolet may preferably be present at an amount of 1.5-5% by weight andmore preferably 2-4% by weight. TABLE 2 CONC. OF GENTIAN VIOLET STAININGCOLOR DEFINITION 1.5% BY WEIGHT ◯˜Δ 2.5% BY WEIGHT ◯ 4.5% BY WEIGHT ◯

Example 4

[0080] Similar process was repeated as in Example 3 except for usingvariable amounts of brilliant green. Sample used was a specimen ofcorneocites obtained from an individual whose skin was in a badcondition in which nuclear cells were observed. The clearness degree ofnuclear staining was determined as in Example 3. The results are shownin Table 3 below. Those results show that brilliant green may preferablybe present at an amount of 0.7-2% by weight and more preferably 0.8-1.5%by weight. TABLE 3 CONC. OF BRILLIANT GREEN STAINING COLOR DEFINITION0.7% BY WEIGHT ◯˜Δ 0.9% BY WEIGHT ◯ 1.5% BY WEIGHT ◯

Example 5

[0081] Similar process was repeated as in Example 1 except for usingvariable amounts of ethanol. Additionally, the presence or absence ofprecipitation (insoluble fraction) of the stain was determined. Theclearness of staining was indicated as: ◯=clear; Δ=less clear; X notclear. The precipitation was indicated as: “present”; “a small amount”;“absent”. The results are shown in Table 4 below. Those results showthat water-soluble organic solvent may preferably be present in theliquid solvent at an amount of 3-40% by weight and more preferably 5-30%by weight. TABLE 4 CONC. OF STAINING COLOR ETHANOL PRECIPITATIONDEFINITION  1% BY WEIGHT SMALL AMOUNT ◯˜Δ 25% BY WEIGHT NONE ◯ 30% BYWEIGHT NONE ◯

Example 6

[0082] Similar process was repeated as in Example 1 except for using a20% ethanol solution as the organic solvent. Clearly stained specimenwas obtained as in Example 1.

Example 7

[0083] Similar process was repeated as in Example 1 except for using a10% acetinitrile solution as the organic solvent. Clearly stainedspecimen was obtained as in Example 1.

Example 8

[0084] Similar process was repeated as in Example 1 except for usingdifferent mounting agents. Color running was indicated as: ◯=no running;Δ=some running; X=significant running. The results are shown in Table 5below. Those results show that oil and fat constituent and/or componentthat is liquid at 1 atm, 25° C. may preferably be used as an mountingagent for preparing horny cell specimen according to the presentinvention. TABLE 5 ENCAPSULATING AGENT COLOR-BLEEDING PHEMETHICON ◯OLEIC ACID OCTYLDO DECYL ◯ GLYCERYL TRIISOOCTANOATE ◯ UV-CURING RESIN XBALSAM/XYLENE SOLUTION X

[0085] According to the present invention, methods for staining hornycell and preparing horny cell specimen can be provided which can clearlystain horny cell during shorter period of time and provide usefulinformation for personal consultation quickly.

[0086] [3] Skin Analysis System

[0087] Embodiment of the skin analysis system according to the presentinvention will hereinafter be described in reference to attacheddrawings though the configuration of the inventive system may not belimited to those described in the embodiment.

[0088]FIG. 4 shows an embodiment of the skin analysis system accordingto the present invention. The skin analysis system shown in FIG. 4 canbe used to provide a customer who visits a cosmetic store with aconsultation including skin analysis (skin evaluation) of the customer(a subject), provide the customer with an opportunity to purchase atrial kit selected based on the results of the evaluation so that he/shecan purchase the cosmetic product(s) contained in the trial kit ifhe/she likes it (them).

[0089] For this purpose, the inventive skin analysis system may comprisea store system or systems 10 which are located in stores, a serversystem 20 which is connected to the store system or systems 10 via anetwork (e.g., using an internet leased line over the internet in thisexample), and an Evaluation personnel computer (Evaluation PC) 30 whichmay be connected to the server system 20 via a network (e.g., a localarea network).

[0090] According to the skin analysis system, the following 6 categoriesmay be analyzed (evaluated): “roughness of the surface of keratinizedlayer (TA); “completeness of keratinized layer”; “texture”; “darkspot/color evenness”; “sensitive skin type”; and “tendency to acne”.

[0091] For this purpose, store system 10 may comprise a horny cellstaining device 11, a microscope 12, an analytical personnel computer 13(Analytical PC), video microscopes 14 and 15, and a consultationpersonnel computer 16 (Consulting PC).

[0092] The horny cell staining device 11 may be used to staincorneocites which have been collected from the surface of the customer'scheek which are then used to analyze the roughness of the surface ofkeratinized layer or the completeness of keratinized layer of thecustomer's skin (of a part of face, e.g., cheek). Corneocites may beobtained by stripping off from the surface of the customer's cheek skinusing an adhesive material. Any known horny cell checker 17 (e.g., atransparent adhesive tape such as cellophane-tape) or surface area disc18 (e.g., an adhesive disc comprising a polyethylene terephthalate plateand adhesive coated thereon) may be used. Corneocites may be stained ina solution of stain in solvent containing a water-miscible organicsolvent. Therefore, corneocites attached to the adhesive material can bestained directly since the substrate of the adhesive material may not bedamaged during the staining step, thereby eliminating the necessity offurther transferring the sample to another material for staining. Stainspreferably include eosin, hematoxylin, gentian violet, brilliant greenand malachite green, which can be used alone or in combination. Amongall, a combination of gentian violet and brilliant green may beparticularly preferable. This is because that cytoplasm can be stainedwith gentian violet to obtain a high contrast between the cytoplasm andthe background thereby clearly visualizing the shape of corneocites, andthat the stained cytoplasm may in turn act as the background so thatnuclei of the corneocites can be clearly stained with brilliant green.The sample is stained in a solution of any of such stains in liquidsolvent containing a water-soluble (water-miscible) organic solvent.Stain may preferably be present in the dyeing solution at an amount of2-7% by weight. For a combination of gentian violet and brilliant green,gentian violet may preferably be present at an amount of 1.5-5% byweight and more preferably 2-4% by weight while brilliant green maypreferably be present at an amount of 0.7-2% by weight and morepreferably at 0.8-1.5% by weight. Examples of water-soluble organicsolvent preferably include: alcohol such as methanol, ethanol,isopropanol or 1,3-butanediol; keton such as acetone or methyl ethylketone; nitrile such as acetinitrile; and ether such as tetrahydrofuran.Among all, alcohol is preferable and ethanol is particularly preferable.Water-soluble organic solvent may preferably be present in the liquidsolvent at an amount of 3-55% by weight and more preferably 5-50% byweight. This is because that a larger amount of organic solvent may notcontribute to reduction in the time required for staining or rather maydegrade the adhesive or substrate of adhesive material while a smalleramount cannot stain a sample clearly. Under those conditions, it maytake 1-5 minutes to stain a sample, which is much shorter than the timerequired for staining according to conventional methods (10-30 minutes)(using 0.5-1% by weight of gentian violet and 0.2-0.5% by weight ofbrilliant green). Further, a very high contrast can be obtained.Besides, the staining method using those conditions can be performed atroom temperature. Staining may be performed by the person in charge instore using horny cell staining apparatus 11.

[0093] Next, the person in charge may prepare a horny cell specimenusing the stained corneocites. A horny cell specimen may be prepared byobtaining a horny cell sample containing corneocites which have beenstripped off from the surface of skin using an adhesive material,staining the horny cell sample and mounting the stained horny cellsample in an oil and fat component and/or composition that is liquid at1 atm, 25° C. Examples of oil and fat constituent and/or compositionthat is liquid at 1 atm, 25° C. may include any oil and fat constituentswhich are conventionally used in cosmetic products and drugs as well ascompositions comprising any combination thereof which are liquid at 1atm, 25° C. Preferable examples may include one or any combination oftwo or more selected from the group consisting of silicone oil, fattyacid triglyceride, ester of higher alcohol and fatty acid, andcarbohydrate which are liquid at 1 atm, 25° C. Generally, mountion in aliquid component may not provide a stability of 1 month or longer.However, this may not be critical for an instant consultation at a storecounter. Such oil base may not cause color running and thus amicroscopic image of a specimen obtained can be processed into a binaryimage (binarization) based on whether the corneocites have been stainedor not to visualize the shape of horny cell accurately. Then, the sizeof horny cell can automatically be computed accurately. Additionalinformation which can be obtained from a specimen prepared by theabove-described method include the presence or absence of nuclear cell,the frequency of nuclear cell appearance, the arrangement regularity ofcorneocites, the shape of horny cell, the roughness of the surface ofkeratinized layer and the like. The specimen can preferably be used todetermine numerical values for those parameters. In summary, horny cellspecimens prepared may preferably be used to determine one or moreselected from the group consisting of the size of horny cell, thepresence or absence of nuclear cell, the frequency of nuclear cellappearance, the arrangement regularity of corneocites, the shape ofhorny cell and the roughness of the surface of keratinized layer.

[0094] Microscope 12 may produce an enlarged image of the horny cellspecimen placed thereon at a desired magnification. In this example, thecorneocites may be magnified 30 times for evaluating the roughness ofthe surface of keratinized layer (TA), and 150 times for evaluating thecompleteness of keratinized layer. Microscope 12 may comprise an imagepickup device such as CCD (charge-coupled device). An enlarged imageproduced by the image pickup device may be then transmitted toAnalytical PC 13 via a signal line.

[0095] Analytical PC 13 may comprise a CPU, a main memory, an auxiliarymemory, a communication control device, an input device (such as akeyboard or a mouse) and an output device (such as a display monitor ora printer). The CPU may load a program obtained from the auxiliarymemory into the main memory and run the program to perform the followingfunctions:

[0096] (A) to receive enlarged images transmitted from microscope 12(output signals from the image pickup device) and generate image datafor these image (horny cell images (TA) and (completeness)), whichcorresponds to a horny cell image data generating means;

[0097] (B) to determine the size of horny cell using the horny cellimage, which corresponds to a measurement means; and

[0098] (C) to transmit the horny cell image data and the horny cell sizedetermined to server system 20 over the Internet, which corresponds totransmitting means.

[0099] Analytical PC 13 may acts as a measurement means to binarize thehorny cell image into black and white (monochromatic image of the hornycell is generated). Therefore, the horny cell image may consist of oneor more black regions (stained horny cell(s)) and white portion(s) (thebackground: unstained region(s)). The number of pixel in the blackregion(s) (horny cell or corneocites) may be determined and the size ofhorny cell may be calculated from the pixel number. The size may bedetermined for two or more corneocites in the horny cell image, and theaverage horny cell size and the number of horny cell to be used todetermine the average size may be recorded.

[0100] Analytical PC 13 may then act as transmitting means to transmitthe image data for each horny cell image as well as information on theaverage size and the number of corneocites measured to server system 20over the Internet.

[0101] Video microscope 14 may produce an enlarged image (X 30) of thesurface of a desired part (e.g., cheek) of customer's face for analyzingthe texture of the customer's skin, and transmit image signals generatedtherefrom to Consulting PC 16.

[0102] Video microscope 15 may produce an enlarged image (X 5) of thesurface of a desired part (e.g., cheek) of customer's face for analyzingthe dark spot/color evenness of the customer's skin, and transmit colorimage signals (RGB signals) generated therefrom to Consulting PC 16.

[0103] Consulting PC 16 may comprise a CPU, a main memory, an auxiliarymemory, a communication control device, an input device (such as akeyboard or a mouse) and an output device (such as a display monitor ora printer). The CPU may load a program obtained from the auxiliarymemory into the main memory and run the program to perform the followingfunctions:

[0104] (a) to receive image signals from video microscope 14 andgenerate image data for them (texture image data), which corresponds toa texture image data generating means;

[0105] (b) to receive color image signals from video microscope 15 andgenerate image data for them (original image data) as well as B channelimage data, i.e., image data consisting of B channel only extracted fromthe original image, which corresponds to a dark spot image generatingmeans;

[0106] (c) to receive information on the customer's attributes (privatedata: name, age, address and the like) and answers to questionnairesabout skin 19 (FAQ's about skin) including skin troubles which have beenentered into Consulting PC 16 using an input device, which correspondsto a reception means;

[0107] (d) to transmit image data for the texture evaluation, originaland B channel images as well as the manually entered data (e.g., thecustomer's attributes, answers to FAQ's and others) to server system 20over the internet, which corresponds to a transmitting means;

[0108] (e) to receive the data of web page(s) containing the results ofskin analysis from server system 20 over the internet, which correspondsto a receiver means; and

[0109] (f) to output the received web page(s) to a monitor (an outputdevice) for display (which corresponds to a display control means) or toa printer for printing.

[0110] Consulting PC 16 may generate a B channel image (blue light imagewith peak at 435.8 nm in principle) as an image to be used for darkspot/color evenness analysis. B channel image is known to be useful fordetecting melanin granules in a horny cell. Conventionally, corneocitescollected from the surface of subject's skin have been stained withsilver nitrate and gentian violet to prepare a specimen which was thenobserved by a microscope to inspect melanin which may cause generationof dark spots (see Japanese Patent Application Laid-Open No.2000-212037). According to the present invention, a B channel image maybe generated using a video microscope 15 to eliminate preparing aspecimen for melanin observation, thereby providing results of dark spotanalysis more quickly than the prior art.

[0111] Horny cell, texture, original and B channel images may beprovided in, for example, JPEG standard data (JPEG data) format, or inany suitable image file format such as GIF or WINDOWS BITMAP.

[0112] Server system 20 may comprise an application server 21 (whichcorresponds to a display data generating device) as the display datagenerating device according to the present invention and a databaseserver (DB server) 22. Application server 21 may comprise a CPU, a mainmemory, an auxiliary memory, a communication control device and otherdevice or devices. The CPU may load a program obtained from theauxiliary memory into the main memory and run the program. In this way,the application server 21 can perform the following functions:

[0113] (I) to receive, from Consulting PC 16 (originally from AnalyticalPC 13), and store the horny cell image data and the information on theaverage size and the number of horny cell measured, which corresponds toa receiving means;

[0114] (II) to receive and store the texture, original and B channelimage data transmitted from Consulting PC 16 as well as the manuallyentered data including the customer's attributes and answers to FAQs,which corresponds to a receiving means;

[0115] (III) to transmit the data and information received fromAnalytical PC 13 and Consulting PC 16 to Evaluation PC 30;

[0116] (IV) to generate data for one or more web pages which containresults of the evaluation obtained based on the data and/or informationon the results of the evaluation transmitted from Evaluation PC 30 andon the data and/or information stored in DB server 22, which correspondsto a data generating means; and

[0117] (V) to transmit the web page data to Consulting PC 16.

[0118] DB server 22 may comprise a CPU, a main memory, an auxiliarymemory, a communication control device and other device or devices. TheCPU may load a program obtained from the auxiliary memory into the mainmemory and run the program. In this way, DB server 22 may perform thefollowing functions:

[0119] (i) to generate and update an aggregate database which is adatabase that contains accumulated data of the results obtained fromskin analysis (evaluation); and

[0120] (ii) to generate and update a customer database which is adatabase that contains accumulated results obtained from skin evaluationfor each customer.

[0121] Before developing the skin analysis system according to thepresent invention, the present inventors performed a certain skinanalysis on hundreds of panelists to examine the roughness of thekeratinized layer, the completeness of corneocites, skin texture, darkspot/color evenness and whether sensitive skin type or not, andpreserved the data obtained. According to the inventive skin analysissystem, DB server 22 stores those data as an aggregate data (mass data)so that those data can be added to the customer's data for comparison.

[0122] Evaluation PC 30 may be located at Analysis Center andmanipulated by an evaluator at the center. Evaluation PC 30 may comprisea CPU, a main memory, an auxiliary memory, a communication controldevice and the like. The CPU may load a program obtained from theauxiliary memory into the main memory and run the program. In this way,Evaluation PC 30 may perform the following functions.

[0123] Evaluation PC 30 may receive image data for horny cell images(TA) and (completeness), a texture image and a B channel image as wellas information on the average size and number of horny cell measuredfrom Application server 21, and display the data and information on adisplay device (a display monitor). The evaluator may evaluate TA andcompleteness by referring to the horny cell images (TA) and(completeness) displayed on the display monitor as well as taking intoconsideration the average size and number of horny cell measured. Theevaluator may also evaluate the texture of skin by referring to thetexture image displayed on the display monitor. Further, the evaluatormay also evaluate dark spot/color evenness by referring to the B channeldata on the display. TA, completeness, texture, dark spot and colorevenness may be evaluated based on, for example, a 10-point scaleevaluation. Evaluation PC 30 may generate information on those resultsof the evaluation which may be then transmitted to application server21.

[0124] Once application server 21 receives the results of the evaluationfrom Evaluation PC 30 it may produce a web page containing thoseresults. For example, application server 21 may produce data for a webpage which may contain the results of the evaluation received fromEvaluation PC 30 (a web page in the detailed analysis zone describedlater). In this step, when data of the customer's past results have beenpreserved in the customer database in DB server 22, those data may beobtained and used to determine the average of the customer's past data(described later) which may be then included in the web page.

[0125] Application server 21 may perform a total evaluation on theresults obtained by current skin analysis of the customer according to apredetermined principles based on the results of the evaluationtransmitted from Evaluation PC 30, determine a total skin score,position and the like (described later), and generate data for a webpage showing them. In this step, an aggregate data on the resultsobtained for those of the same generation may be obtained from theaggregate data base in DB server 22 and used to create a web page sothat the results of the evaluation of the customer can be compared tothe aggregate data. When the customer' previous data are preserved inthe customer database in DB server 22, the data may be obtained andincluded in the web page as the previous data (described later).

[0126] Moreover, application server 21 may determine advice on skincare(skincare advice), product or products recommended to be used forskincare (cosmetic products) and a trial kit based on the results of theevaluation, and produce a web page which presents those information.

[0127] Data materials to be used for producing the above-described webpage (e.g., HTTP file, XML file, image/dynamic image/audio file or thelike) may have already been prepared in the auxiliary memory ofapplication server 21. Application server 21 (or the CPU thereof) mayproduce a web page using the data components. In this step, the hornycell, texture, original and B channel images and/or other data receivedfrom Consulting PC 16 may be used as components of the web page. Variousweb pages which each present different cosmetic products or trial kitsmay be prepared in the auxiliary memory of application server 21 (or DBserver 22), and suitable one may be selected among these pages accordingto the recommended skincare procedure determined based on the results ofthe evaluation for the customer.

[0128] In this way, application server 21 may construct a web siteconsisting of a plurality of web pages showing the results of skinanalysis (results of the evaluation) and transmit the data for those webpages to Consulting PC 16 over the internet.

[0129] The results of the evaluation of the current skin analysisdisplayed on the web pages may be accumulated in the customer databaseof DB server 22 as well as in the aggregate data base as a part ofaggregate data.

[0130] Application server 21 may supply the results of skin analysisobtained for the customer in a web system format to Consulting PC 16. Insummary, a plurality of web pages containing information such as skinanalysis results may be transmitted to Consulting PC 16 over theInternet. Once Consulting PC 16 receives those web pages, a web browser(a display control means) installed in Consulting PC 16 may display eachpage on the screen monitor. The customer can receive the skin analysisresults, advice on skin treatment and explanation about cosmeticproducts which are recommended to the customer by referring to the webpages.

[0131]FIG. 5 is a block diagram of a web site (referred to as “advicesite”) consisting of a plurality of web pages which will be provided toa customer, showing a plurality of different web pages and therelationship of the linkages therebetween. In FIG. 5, the advice sitemay comprise an entertainment (introduction) zone, a detailed analysis(laboratory) zone, and a product explanation (cosmetic information)zone.

[0132] The entertainment zone may consist of web pages each titled “1.total skin score (total evaluation)”, “2. general chart” and “3. skinpositioning”. The detailed analysis zone may consist of web pages titled“4. roughness of the surface of keratinized layer”, “5. completeness ofkeratinized layer”, “6. texture”, “7. dark spot/color evenness” and “8.sensitive type”. The product explanation zone may consist of web pagestitled “9. skincare advice” and “10. trial kit”. The advice site mayfurther contain web pages titled “11. skin biorhythm” and “12. utility”.

[0133]FIG. 6 is an explanatory view of a web page representation. InFIG. 6, specifications of the web page representation are provided inthe box (dashed line) (similarly in FIGS. 7-22). As shown in FIG. 6, amenu list area showing the list of web page titles (analysis menu) isprovided on the left side of the page screen. Indexes for web pages Nos.1-12 (shown in FIG. 5) are provided in the menu list area. Each indexmay function as the icon for shortcut to the designated web page and canbe selected by using an input device (a keyboard or a mouse). Once aparticular index is selected, the selected index may be highlighted andindicates the current location on the advice site. The menu list areamay be permanently displayed at the advice site. A main display area maybe provided on the right side of the menu list area. Each of web pagesNos. 1-12 may be alternately displayed in the main display area.

[0134]FIG. 7 shows one example of the top (title) web page of the advicesite. In this page, the title of the advice site, customer's name,customer's identification information (ID), and date of test are shownin the main display area. Each of customer's name, ID and date of testcolumns may also function as an entry space for those information.

[0135]FIG. 8 shows one example of web page titled “1. total skin score(total evaluation)”. Customer's name, his/her total skin score (thetotal score obtained by evaluation of horny cell) and skin positioning(which indicates the ranking of the results of the evaluation). “Nobleskin” is one of the zone names indicating horny cell rankings.

[0136]FIG. 9 shows one example of a first web.page titled “2. generalchart”. In the first web page, the main display area may provide a radarchart showing the tendency of the customer's skin. The radar chart mayshow a 10-point scale evaluation for each of 6 categories: “ability tokeep moisture”; “oiliness (sebum)”; “tendency to acne”; “dark-spot/colorevenness”; “wrinkle/texture”; and “sensitive skin type”. Each categorycolumn located at the end of an extended line of each axis of the radarchart may function as a shortcut icon linked to the associated page.Particularly, when one category column is clicked by a mouse, the screenmay jump to the corresponding web page (any one of web pages Nos. 4-8)in the evaluation analysis zone. The main display area may also containcheck-boxes “Current data”, “Previous data”, “Average of past data” and“Average for the same generation” at its lower right. The representationof the radar chart may be selectively changed by checking these checkboxes. “Current data” indicates the results obtained by the current skinanalysis of the customer, “Previous data” indicates the result from theprevious skin analysis, and “Average of past data” indicates the averageof the results obtained for each category in the past skin analyses. The“same generation” refers to people of ages of a predetermined range withrespect to the customer's age (for example, the customer's age ±1 or 2,and, in this example, the customer's age ±1). Accordingly, “Average ofthe same generation” indicates the average of results obtained for eachcategory obtained from the mass data (aggregate data) based on such skinanalyses performed on a large number of people of the same generation.The main display area may also provide the number of the aggregate datasample used to determine the “average of the same generation”. “Yourskin type” and “Comparison with the same generation (ages 24-26)” iconsare provided under the check boxes. “Your skin type” icon may be aswitch icon to the first page from the second page (i.e., make the firstpage active) while “Comparison with the same generation (ages 24-26)”icon maybe a switch icon to the second page from the first page (i.e.,make the second page active). One of these switch icons which iscurrently active may be highlighted (i.e., displayed in a deeper color),thereby indicating which page is currently active. In the first page,“Your skin type” icon may be highlighted. Further, problem items checkedby the customer in his/her answers to the FAQ's (subjective symptomscited directly from the answer) may be displayed as “Your problem (s)”in the lower left of the main display area. In this way, the customercan instantly recognize his/her positioning relative to the mass data ordetermine if his/her skin conditions have been improved by comparinghis/her “current data” with “previous data”, “average of past data” and“average for the same generation” displayed on the radar chart.

[0137]FIG. 10 shows one example of a second web page titled “2. generalchart”. In the second page, the main display area may illustrate a graphshowing the comparison between the customer's current data with theaverage data for the same generation. In this graph, the center axisindicates the average for the same generation, and the customer's dataon each of 6 categories obtained in current skin analysis are shown inrelation to the average. Particularly, a bar which extends into onedirection (e.g., into the right in FIG. 10) may indicate a resultsuperior to the average while a bar which extends into the otherdirection (e.g., into the left in FIG. 10) indicates a result inferiorto the average in the bar graph. Results of the evaluation for thecustomer may be expressed in a 10-point scale (5 scales for superior andinferior each). In the main display area on the second page, symbols maybe provided in the vicinity of the respective categories so that whetherthe results for the customer are superior or inferior to the average forthe same generation can be recognized easily. In this example,categories for which superior results have been obtained may be markedwith “sun (fine)” mark, relatively inferior with “cloud (cloudy)” mark,and much inferior with “umbrella (rainy)” mark. Under the graph, therange of age (in this example, ages 24-26 with respect to age 25, thecustomer's age) and the number of samples from which the average hasbeen obtained may be provided. Below those information, switch icons areprovided which are similar to those in the first page so that the secondpage can be reversibly switched to and from the first page. In thesecond page, the “comparison with the same generation” icon may behighlighted when compared to the “your tendency”.

[0138]FIG. 11 shows one example of a web page titled “3. skinpositioning”. In the main display area on this page may be provided atotal skin score and a skin position determined in the present skinanalysis for the customer as well as his/her average data of the pastskin analysis. A “total skin score” is the sum of the points determinedfor the respective 6 categories in a skin analysis and indicates thetotal result of the customer's skin horny cell. A “skin positioning” mayindicate the location (rank) where the customer's total result of theskin analysis (i.e., skin condition) is located in the distributions ofthe results for the same generation, which may be represented in a10-position scale (divided in 10 groups) with 5 positions (2 scales foreach position), each titled, for example, “ragged”, “dangerous”,“normal”, “noble” and “queen” (sequentially from the worst to the best)in this embodiment. Past average score may comprise the average of atleast 2 skin analyses which have been performed on the customer in thepast. In the main display area, a skin positioning graph may also beprovided which indicates the distribution of skin positioning obtainedfor panelists of the same generation. The customer's skin positioning(zone) maybe provided in the skin positioning graph. Such a skinpositioning graph may be prepared and displayed with the reference(average) data of the corresponding month (i.e., such reference averagedata may be prepared for each month) since the skin condition may changecorresponding to seasonal change. Therefore, the panelist's dataobtained during the same month as the date of the current analysis maybe extracted from the aggregate data and used to prepare the skinpositioning graph.

[0139]FIG. 12 shows an example of a web page titled “4. roughness of thesurface of keratinized layer (the ability of keratinized layer to keepmoisture)”. In the main display area on this page may be provided ascore (represented in a 5-point scale) determined on the roughness ofthe surface of keratinized layer observed in the customer as well as theaverage of his/her past scores. A microscopic photograph (×30) of thecustomer's keratinized layer may also be provided in the main displayarea. The microscopic photograph may be an image reproduced from theimage data (JPEG data) for an enlarged image of the customer'scorneocites which has been obtained by microscope 12 and processed inAnalytical PC 13. Information on how to evaluate the roughness or thelike may be provided in the vicinity of the microscopic photograph. Thecustomer's score and a standard sample photograph may also be providedin the main display area.

[0140]FIG. 13 shows one example of a web page titled “5. completeness ofkeratinized layer”. In the main display area on this page may beprovided a score (represented in a 5-point scale) determined on thecompleteness of the customer's keratinized layer as well as the averageof his/her past scores. A microscopic photograph (×150) of thecustomer's keratinized layer may also be provided in the main displayarea. The microscopic photograph may be an image reproduced from theimage data (e.g., JPEG data) for an enlarged image of the customer'scorneocites which has been obtained by microscope 12 and processed inAnalytical PC 13. Information on how to evaluate the completeness ofkeratinized layer or the like may be provided in the vicinity of themicroscopic photograph. The customer's score and a standard samplephotograph may also be provided in the main display area. Further, thehorny cell size of the customer may also be provided.

[0141]FIG. 14 shows one example of a web page titled “6. texture”. Inthe main display area on this page may be provided a score (representedin a 5-point scale) determined on the texture of the customer's skin(texture condition) observed in the present analysis as well as theaverage of his/her past scores. An enlarged photograph (×30) of thecustomer's skin surface showing its texture may also be provided in themain display area. This enlarged photograph may be an image reproducedfrom the image data (e.g., JPEG data) for an image of the surface of thecustomer's skin obtained by video microscope 14 and processed inConsulting PC 16. Information on how evaluate the texture condition orthe like may be provided in the vicinity of the microscopic photograph.The customer's score and a standard sample photograph may also beprovided in the main display area.

[0142]FIG. 15 shows one example of a web page titled “7. dark spot/colorevenness”. In the main display area on this page may be provided a score(represented in a 5-point scale) determined on the dark spot/colorevenness observed in the current analysis as well as the average ofhis/her past scores. An RGB color photographic image (original image:×5), which is an enlarged image of the customer's skin surface and aphotographic image (B channel image) comprising B channel extracted fromthe RGB color image may also be provided in the main display area. Theoriginal and B channel images may be images reproduced from image data(e.g., JPEG data) for an enlarged image of the surface of the customer'sskin which have been obtained by video microscope 15 and processed inConsulting PC 16. Information on how to evaluate dark spot/colorevenness or the like may be provided in the vicinity of the original andB channel images. The customer's score and a standard sample photographmay also be provided in the main display area.

[0143]FIG. 16 shows one example of a web page titled “8. sensitive skintype”. In the main display area on this page may be provided 5categories as indicators indicative of sensitive skin type, including “aprotective ability of keratinized layer”, “resistance against chemicalstimulation”, “resistance against physical stimulation”, “resistance toUV-light”, and “adaptivity to environmental change” as well as a graphwhich show the results on the respective categories obtained for thecustomer. In the graph, a higher (longer) bar means a better result. InFIG. 13, a longer the bar which extends into the right directionindicates a better result. Average for the same generation may beindicated by dotted line for each category in FIG. 13. Particularlyimportant category or categories for skincare for the customer may bemarked with a symbol indicating as such in the vicinity of the categoryor categories. Further, each category title provided in the main displayarea may act as the shortcut icon to the corresponding explanatory pagewhich explains the term and the reason on which the results of theevaluation is based. Jumping to the explanatory page of any desiredcategory can be executed by clicking on a jumping icon by using a mouse.

[0144]FIG. 17 shows one example of a web page titled “9. skincareadvice”. In the main display area on this page may be provided importantpoint(s) on skincare (treatment) recommended for the customer (in. thisexample, “whitening” care). The mutual relationship of 2 parameters(indicators) “an ability to keep moisture” and “sebum” may beschematically shown by a matrix consisting of 9 regions (skincarezones). In this matrix, the zone which corresponds to the customer'sresult may be highlighted. A flow chart showing an appropriate skincareprocedure (skincare line) may also be provided in the main display area.The name of item is indicated in a block representing for each step.Each of those blocks may act as a shortcut icon to the introduction pageof cosmetic product(s) (which correspond(s) to the item name indicatedin the block) which is(are) recommended for use in the step.

[0145]FIG. 18 shows one example of a web page which introduces cosmeticproducts to be used in “lotion” and “moisture essence” steps. This pagecorresponds to the page displayed after jumping executed by clicking oneither “lotion” or “moisture essence” icon in the flow chart shown inFIG. 17. In this page, particular cosmetic products recommended for thecustomer to used in “lotion” and “moisture essence” steps as well as theproperties, volumes, prices and water/oil mark of the products, and theskincare zones covered by the respective products may be presented.

[0146]FIG. 19 shows one example of a web page which introduces cosmeticproducts to be used in “cleansing” and “(facial) wash” steps. This pagecorresponds to the page displayed after jumping executed by clicking oneither “cleansing” and “facial washing” icons in the flow chart shown inFIG. 17. In this page, particular cosmetic products recommended for thecustomer to use in “cleansing” and “facial wash” steps as well as theproperties, volumes, prices and water/oil mark of the products, and theskincare zones covered by the respective products may be presented.

[0147]FIG. 20 shows one example of a web page which introduces cosmeticproducts to be used as “special item(s)”. This page is an introductionpage of cosmetic products which are recommended as special items in theflow chart shown in FIG. 17. Those special items correspond to “moisturecoat”, “pharmaceutical whitening spots” and “clay pack” in the flowchart shown in FIG. 17. Therefore, this page may be displayed afterjumping executed by clicking on either one of these item blocks. In thispage, particular cosmetic products recommended for the customer to usein “moisture coat”, “pharmaceutical whitening spots”, “clay packcleansing”, and “additional items” steps as well as the properties,volumes, prices and the like thereof may be provided.

[0148]FIG. 21 shows one example of a web page titled “10. trial kit”.This page may introduce a trial kit which contains a combination ofcosmetic products recommended for the customer which have been selectedbased on the results of skin analysis. In this embodiment shown in FIG.21, information on the trial kit recommended may be displayed whichinclude the product names, properties, volumes, water/oil mark or otherinformation of the cosmetic products corresponding to “lotion”,“moisture essence” and “facial wash” as well as the price of the trialkit.

[0149]FIG. 22 shows one example of a web page titled “11. skinbiorhythm”. In the main display area on this page may be provided atime-course graph showing the change in the customer's skin conditionbased on the 6 categories in skin analysis during one year and, forcomparison, a corresponding reference graph showing the average of theresults obtained for the same generation. Check boxes for the respective6 categories may also be provided in the main display area. When one ofthese 6 categories is checked, two graphs each representing thecustomer's results on the checked category and the corresponding averageresults for the same generation regarding that category may bedisplayed. It may be required that the customer take a skin testperiodically to prepare such a reference graph.

[0150] According to the above-described skin analysis system, thefollowing operation may be performed in store. When a customer takes askin test in store using store system 10, the customer may have toremove make-up and then his/her corneocites may be collected and/or anenlarged image of the surface of a portion (cheek) of his/her face skinmay be obtained.

[0151] Next, the counselor (adviser) at the store counter may provide aquestionnaire to the customer. The customer may give answers to thequestionnaire including the customer's attributes and answers to theFAQ's about skin 19 (including skin problems) These answers may be inputin Consulting PC 16. The input information may be automaticallytransmitted to server system 20.

[0152] Then, the counselor may obtain enlarged images (×30 and ×5) ofthe surface of the customer's cheek using video microscopes 14 and 15.These images may be transmitted to Consulting PC 16 which may thenproduce image data of texture, original and B channel images based onthese images and then transmit those data to server system 20automatically.

[0153] Next, the counselor may inspect the customer's corneocites.Particularly, horny cell checker 17 and/or size disc 18 may be broughtinto contact with his/her cheek to strip corneocites off therefrom tocollect a horny cell sample which is then stained in horny cell stainingdevice 11 to prepare a horny cell specimen which may in turn be placedon microscope 12. Then, enlarged images of corneocites obtained atdesired magnifications (×30 and ×150) by microscope 12 may be thentransmitted to Analytical PC 13 where the average size and number ofhorny cell. observed may be determined. Those data may be thentransmitted together with the horny cell images to server system 20automatically.

[0154] In a short time, Consulting PC 16 may receive web page data fromserver system 20 and display the web page on its display monitor. Thecounselor and/or the customer can see any one of web pages 1-12 asdesired by using the input device connected to Consulting PC 16. In thisway, the customer can receive his/her total skin score, general chart,skin positioning, results of detailed analysis or evaluation as well asan explanation of products.

[0155] The counselor may give additional advice to the customer whileshowing the web pages to the customer. The customer may purchase a trialkit selected according to the advice provided on the web page and cantry it there. After using the trial kit, the customer can purchase afull size product or products of any items contained in the trial kit ifhe/she likes it/them since the volume of products in such a trial kit isvery small. Alternatively, the customer can purchase any product orproducts introduced in the skincare procedure recommended for thecustomer without using a trial kit containing it/them.

[0156] The time required from the cleansing to provision of the resultsof analysis (i.e., web pages from page 1) maybe reduced to 30 minutes byemploying the above-described methods for staining horny cell andpreparing specimen as well as production of B channel image. Therefore,the customer can receive the results of skin analysis, advice on skincare procedure determined according to those results and an explanationof the cosmetic product(s) to be used in the recommended skincareprocedure on the day he/she visits the store. With such convenience,customers may have more opportunities to purchase cosmetic products, andtherefore the sales of cosmetic products may be promoted.

[0157] The customer's data on his/her results of the evaluation may beprovided to the customer together with reference data prepared fromanalysis performed on those of the customer's age ±1 or 2 (average datafor the same generation) such that those data can be compared to eachother (see FIGS. 9, 11 and 16). Alternatively, the customer's resultsmay be provided to him/her which are represented in relation to resultsfor the same generation (see FIG. 10). Therefore, the customer can knowhis/her skin condition more specifically.

[0158] The results of the evaluation for the customer may be then storedin DB server 22 every time so that the previous data may be displayedtogether with the current results of the evaluation (see FIG. 9). Thismay allow the customer to know if his/her skin condition has beenimproved or not. The customer can also know much more detail abouthis/her skin condition by referring to the average of his/her past datafor each category. Further, from his/her past data, a time-coursepattern of change in the customer's skin condition may be provided as askin biorhythm in comparison with the average pattern for the samegeneration (a standard pattern), by which the customer can know his/herskin condition more specifically.

[0159] Server system 20 may produce web pages containing those resultsof the evaluation and results of the evaluation for each customer may bestored in server system 20. Therefore, total management of the resultsof the evaluation for all the customers may be possible even when aplurality of store systems 10 are located at different stores.

[0160] The skin analysis system according to the present invention mayallow for a quick and clear staining of corneocites as well as quickacquisition of information useful for consultation.

1-4. (Canceled)
 5. A method for preparing a corneocites specimen,comprising: obtaining a sample containing corneocites by stripping askin surface using an adhesive material; staining the sample; andmounting the stained sample onto an oil and fat constituent and/orcomposition which is liquid at 1 atm, 25° C.
 6. The method according toclaim 5, wherein the oil and fat constituent and/or compositioncomprises one or more components selected from the group consisting ofsilicone oil, fatty acid triglyceride, ester of higher alcohol and fattyacid, and carbohydrate.
 7. The method according to claim 5, wherein thesample containing corneocites is stained in a stain solution in solventcontaining a water-miscible organic solvent.
 8. The method according toclaim 5, wherein the corneocites specimen is used to determine ormeasure one or more property selected from the group consisting of sizeof a horny cell, presence or absence of a nuclear cell, frequency ofnuclear cell appearance, arrangement regularity of corneocites, shape ofa horny cell, and roughness of a surface of keratinized layer. 9-17.(Canceled)